miR-375在AGEs介导糖尿病血管损伤细胞中的生物学功能

目的：探讨miR-375在血管损伤细胞中的表达及生物学功能。方法：利用基因克隆技术构建miR-375表达载体;然后将miR-375表达质粒转染至血管损伤细胞中,同时分别设立Huvec12对照组,血管损伤细胞组,血管损伤抑制组,Huvec12转染miR-375组。24h后收集细胞,在mRNA和蛋白水平检测Mtpn、NFκB、profilin1、sICAM1的表达,经荧光染色观察细胞F-actin的变化,再用流式细胞仪检测细胞凋亡。结果：血管损伤细胞中过表达miR-375后,在mRNA和蛋白水平靶基因Mtpn下降,NFκB的表达活性下降,使糖尿病血管病变的标志profilin1下调;F-actin表达恢复;细胞粘附因子（sICAM1）表达下降,细胞凋亡减少。结论：初步证明miR-375可以抑制AGEs介导的糖尿病血管细胞损伤的发生,可能成为糖尿病血管损伤并发症基因治疗的靶点。

The Biological Function of miR-375 in Vascular Cells Injury Induced by AGEs-mediated Diabetes

Objective： To investigate the expression and biological function of miR-375 in injured vascular cells. Methods： Cloning technology was used to construct miR-375 expression vector and subsequently mir-375 expression plasmid was transfected into injured vascular cells. Meanwhile, we set for further study： Huvec12 as control, injured vascular cells, vascular cells after injury inhibition and cells with miR-375 overexpression. Cells were harvested 24 hours after treatments. The expression of Mtpn, NFκB, profilin1 and sICAM1 were measured in both mRNA and protein level. F-actin was examined via fluorescence staining. Cell apoptosis was assessed by flow cytometry. Results： In injured vascular cells with miR-375 overexpression, the mRNA and protein expression of targeted gene Mtpn declined. It was also observed the reduction of NFκB activity, downregulation of the diabetic vascular injury marker profilin1, restoration of F-actin expression, a decrease of sICAM1 expression and reduced cell apoptosis. Conclusions： miR-375 may inhibit the injuries of vascular cells caused by AGEs-mediated diabetes and may become a new target for gene therapy to treat vascular complications caused by diabetes.